Introduction 

Timely in-process monitoring of critical components in culture media is vital for optimal cell growth, quality, and yield in biotherapeutics process optimization. Herein we describe a high throughput LC-MS based method with the widest target compound coverage for the analysis of cell culture media nutrient and metabolites currently available.  The method consists of a 9 min run time, a 220+ targeted compound library with user-friendly data review workflows and access to multivariate data analytics. Performance metrics for qualitative and quantitative determination of cell culture media nutrient and metabolite analysis in commercial cell culture media and spent media are presented.

Methods 

Commercial media solutions were purchased from Millipore Sigma. Spent media samples from a fed-batch bioprocess experiment was carried out using NISTCHO cell line (nist.gov) to produce cNISTmAb product. The samples were diluted from 1:100 to 1:400 using 0.1%FA and subjected to LC-MS analysis. LC-MS instrument used was a BioAccord HRMS system using an ACQUITY Premier LC system.  I Waters_connect was used for data acquisition and processing.

Preliminary Data 

A high throughput 9 min method for cell culture nutrient and metabolite analysis was developed with expanded 220+ compound library. The 9 min method run time represents 50 percent reduction in time compared to the previously published 20 min methods while maintaining the same target compound coverage. The method was employed for spent media analysis of harvested samples from the NISTCHO host producing cNISTmAb as part of overall assessment including titer measurement. Spent media solutions under different glucose feeding conditions were sampled during a 14-day incubation and analyzed using both the 9 min fast method and the 20 min method. Results showed excellent correlation where same trends were obtained using either method, indicating high quality data obtained in the 20-min method is maintained using the rapid 9 min method. Result also show a good correlation of glucose data obtained using either LC-MS method or the Nova Flex2 media analyzer. In addition, the method’s broad applicability was assessed by analyzing representative commercial media solutions, including Waters amino acids cell culture standard solution, DMEM, IMDM, CHO Fed-batch media, HEK293 viral vector media, and microbial growth media. For all media samples, the 9 min method produced the same detection results as for the 20 min method. Excellent accuracy (<15%) and reproducibility (<10%) were obtained using amino acid as standards.